IDENTIFICAÇÃO MOLECULAR DE OSTRAS NATIVAS NA COSTA DO MARANHÃO, BRASIL
DOI:
https://doi.org/10.20950/1678-2305.2018.44.4.377Palavras-chave:
DNA mitocondrial, COI, identificação de espéciesResumo
As ostras são encontradas ao longo de toda a costa do Brasil. Os fenótipos das espécies variam consideravelmente de acordo com as características do habitat. O presente estudo investigou a existência de diferentes espécies de ostra do gênero Crassostrea no litoral do Maranhão, utilizando a técnica de PCR Multiplex e DNA barcoding. Os resultados da PCR Multiplex revelaram duas bandas distintas, características das espécies C. gasar e C. rhizophorae num total de 135 amostras analisadas. No sequenciamento do gene COI de 98 amostras obteve-se fragmentos de 695 pb e 15 haplótipos para C. gasar e de 640 pb e oito haplótipos para C. rhizophorae. A árvore haplótipica agrupou fortemente as duas espécies em clados diferentes com 100% de bootstrap. Divergências intraespecíficas foram de 0,2% para ambas, e a interespecífica de 23,6%. A similaridade das sequencias deste estudo com sequências presentes no BoldSystems variou de 97,01 a 98,37 para C. rhizophorae e de 97,55 a 99,84 tanto para C. gasar como para C. brasiliana, reforçando uma problemática na taxonomia do grupo, que apontam para a sinonimia dessas espécies. Portanto, o DNA barcoding permitiu identificar as amostras coletadas e confirmou a existência de duas espécies de ostraReferências
BONDIOLI, A.C.V.; MARQUES, R.C.; TOLEDO, L.F.A.; BARBIERI, E. 2017 PCR-RFLP for identification of the pearl oyster Pinctada imbricate from Brazil and Venezuela. Boletim do Instituto de Pesca, 43(3): 459-463.
CARVALHO, D. D.; SEERIG, A.; MELO, D. D.; SOUSA, A. B.; PIMENTA, D.; OLIVEIRA, D. A. A. 2008 Identificação molecular de peixes: o caso do Surubim (Pseudoplatystoma spp.). Revista Brasileira de Reprodução Animal, 32(4): 215í 219.
CAVALEIRO, N. P.; SOLÉ-CAVA, A. M.; LAZOSKI, C.; CUNHA, H. A. 2013 Polymorphic microsatellite loci for two Atlantic oyster species: Crassostrea rhizophorae and C. gasar. Molecular biology reports 40(12): 7039-7043.
CAVALLI, R. O.; FERREIRA, J. F. 2010 O futuro da pesca da aquicultura marinha no Brasil: a maricultura. Ciência e Cultura, 629(3): 38-39.
CHRISTO, S. W.; ABSHER, T. M. 2006 Reproductive period of Crassostrea rhizophorae (Guilding, 1828) and Crassostrea brasiliana (Lamark, 1819) (Bivalvia:Ostreidae) in Guaratuba bay, Paraná, Brazil. Journal of Coastal Research, 2(39): 1215-1218.
CYWINSKA, A.; HUNTER, F. F.; HEBERT, P. D. 2006 Identifying Canadian mosquito species through DNA barcodes. Medical and veterinary entomology, 20(4): 413-424.
FELSENSTEIN, J. 1985 Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39(4): 783-791.
FOLMER, O.; BLACK, M.; HOEH, W.; LUTZ, R. VRIJENHOEK, R. 1994 DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3(5): 294í 299.
GALVÃO, M.S.; PEREIRA, O.M.; HILSDORF, A.W. 2013 Molecular identification and distribution of mangrove oysters (Crassostrea) in an estuarine ecosystem in Southeast Brazil: implications for aquaculture and fisheries management. Aquaculture Research, 44(10): 1589-1601.
GALVÃO, M.S.N.; ALVES, P.M.F.; HILSDORF, A.W.S. 2017 First record of the Saccostrea oyster in Bertioga, São Paulo, Brazil. Boletim do Instituto de Pesca, 43(4): 638-645.
GARDUNHO, D.C.L.; GOMES, C.P.; TAGLIARO, C.H.; BEASLEY, C.R. 2012 Settlement of an unidentified oyster (Crassostrea) and other epibenthos on plastic substrates at a northern Brazilian mangrove island. Brazilian Journal of Aquatic Science and Technology, 16(1): 41-51.
HALL, T.A. 1999 BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95 âÂ"ž 98 âÂ"ž NT. Nucleic Acids Symposium Series, 41(41): 95í 98.
HEBERT, P. D.; RATNASINGHAM, S.; DE WAARD, J. R. 2003 Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London B: Biological Sciences, 270 (Suppl 1): S96-S99.
IGNACIO, B.L.; ABSHER, T.M.; LAZOSKI, C.; SOLE´-CAVA, A.M. 2000 Genetic evidence of the presence of two species of Crassostrea (Bivalvia: Ostreidae) on the coast of Brazil. Marine Biology, 136(6): 987í 991.
LAPEGUE, S.; BOUTET, I.; LEITÃO, A.; HEURTEBISI, S.; GARCIA, P.; THIRIOTUIE-VREUX, C.; BOUDRY, P. 2002 Trans-Atlantic distribution of a mangrove oyster species revealed by 16S mtDNA and karyological analyses. Biology Bull, 202(3): 232-242.
LAZOSKI, C.; GUSMÃO, J.; BOUDRY, P.; SOLE´-CAVA, A.M. 2011 Phylogeny and phylogeography of Atlantic oyster species: evolutionary history, limited genetic connectivity and isolation by distance. Marine Ecology Progress Series, 426: 197í 212.
LIBRADO, P.; ROZAS, J. 2009 DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25(11): 1451-1452.
LUDWIG, S.; PATELLA, R.; STOIEV, S.; CASTILHO-WESTPHAL, G.; GIROTTO, M. V.; OSTRENSKY, A. 2011 A molecular method to detect and identify the native species of southwestern Atlantic Crassostrea (Mollusca: Ostreidae). Zoologia (Curitiba), 28(4): 420-426.
MEDRANO, J. F.; AASEN, E.; SHARROW, L. 1990 DNA extraction from nucleated red blood cells. Biotechniques, 8(1): 43.
MELO, A. G. C. D.; VARELA, E. S.; BEASLEY, C. R.; SCHNEIDER, H.; SAMPAIO, I.; GAFFNEY, P. M.; TAGLIARO, C. H. 2010 Molecular identification, phylogeny and geographic distribution of Brazilian mangrove oysters (Crassostrea). Genetics and Molecular Biology, 33(3): 564-572.
MELO, M.A.D.; SILVA, A.R.B.; BEASLEY, C.R.; TAGLIARO, C.H. 2013 Multiplex species-specific PCR identification of native and non-native oysters (Crassostrea) in Brazil: a useful tool for application in oyster culture and stock management. Aquaculture International, 21(06): 1325í 1332.
PIE, M. R.; RIBEIRO, R. O.; BOEGER, W. A.; OSTRENSKY, A.; FALLEIROS, R. M.; ANGELO, L. 2006 A simple PCRâ€ÂRFLP method for the discrimination of native and introduced oyster species (Crassostrea brasiliana, C. rhizophorae and C. gigas; Bivalvia: Ostreidae) cultured in Southern Brazil. Aquaculture Research, 37(15): 1598-1600.
REECE, K. S.; CORDES, J. F.; STUBBS, J. B.; HUDSON, K. L.; FRANCIS, E. A. 2008 Molecular phylogenies help resolve taxonomic confusion with Asian Crassostrea oyster species. Marine Biology, 153(4): 709í 721.
RIOS, E. C. 1994 Sea shells of Brazil. 2ª ed. Rio Grande: Fundação Universidade do Rio Grande. 492p.
SAITOU, N.; NEI, M. 1987 The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4(4): 406í 425.
SANGER, F.; NICKLEN, S.; COULSON, A. R. 1977 DNA sequencing with chain-terminating inhibitors. Proceedings of the national academy of sciences, 74(12): 5463-5467.
SIMONE, L. R. L.; DO AMARAL, V. S. 2014 Revision of genus Crassostrea (Bivalvia: Ostreidae) of Brazil. Journal of the Marine Biological Association of the United Kingdom, 94(4): 811-836.
SOLÉ-CAVA, A. M. 2008 Códigos de barra de DNA: o rabo que abana o cachorro. Ciência Hoje, 41(245): 65-67.
SOLÉ-CAVA, A. M.; Wí–RHEIDE, G. 2007 The perils and merits (or the Good, the Bad and the Ugly) of DNA barcoding of sponges í a controversial discussion. In: CUSTÓDIO, M.R.; LÔBO-HAJDU G.; HAJDU, E.; MURICY G. (ed.) Porifera research: biodiversity, innovation and sustainability. Série Livros (28). Rio de Janeiro: Museu Nacional. p. 603í 612.
TAMURA, K.; STECHER, G.; PETERSON, D.; FILIPSKI, A.; KUMAR, S. 2013 MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular biology and evolution, 30(12): 2725-2729.
THOMPSON, J. D.; HIGGINS, D. G.; GIBSON, T. J. 1994 Clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic acids research, 22(22): 4673-4680.
VARELA, E.S; BEASLEY, C.R.; SCHNEIDER, H.; SAMPAIO, I.; MARQUES-SILVA, N.S.; TAGLIARO, C.H. 2007 Molecular phylogeny of mangrove oysters (Crassostrea) from Brazil. Journal of molluscan studies, 73(3): 229-234.
WAKAMATSU, T. 1973 A ostra de Cananéia e o seu cultivo. São Paulo: Instituto Oceanográfico, Universidade de São Paulo. 141 p.
CARVALHO, D. D.; SEERIG, A.; MELO, D. D.; SOUSA, A. B.; PIMENTA, D.; OLIVEIRA, D. A. A. 2008 Identificação molecular de peixes: o caso do Surubim (Pseudoplatystoma spp.). Revista Brasileira de Reprodução Animal, 32(4): 215í 219.
CAVALEIRO, N. P.; SOLÉ-CAVA, A. M.; LAZOSKI, C.; CUNHA, H. A. 2013 Polymorphic microsatellite loci for two Atlantic oyster species: Crassostrea rhizophorae and C. gasar. Molecular biology reports 40(12): 7039-7043.
CAVALLI, R. O.; FERREIRA, J. F. 2010 O futuro da pesca da aquicultura marinha no Brasil: a maricultura. Ciência e Cultura, 629(3): 38-39.
CHRISTO, S. W.; ABSHER, T. M. 2006 Reproductive period of Crassostrea rhizophorae (Guilding, 1828) and Crassostrea brasiliana (Lamark, 1819) (Bivalvia:Ostreidae) in Guaratuba bay, Paraná, Brazil. Journal of Coastal Research, 2(39): 1215-1218.
CYWINSKA, A.; HUNTER, F. F.; HEBERT, P. D. 2006 Identifying Canadian mosquito species through DNA barcodes. Medical and veterinary entomology, 20(4): 413-424.
FELSENSTEIN, J. 1985 Confidence limits on phylogenies: an approach using the bootstrap. Evolution, 39(4): 783-791.
FOLMER, O.; BLACK, M.; HOEH, W.; LUTZ, R. VRIJENHOEK, R. 1994 DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology, 3(5): 294í 299.
GALVÃO, M.S.; PEREIRA, O.M.; HILSDORF, A.W. 2013 Molecular identification and distribution of mangrove oysters (Crassostrea) in an estuarine ecosystem in Southeast Brazil: implications for aquaculture and fisheries management. Aquaculture Research, 44(10): 1589-1601.
GALVÃO, M.S.N.; ALVES, P.M.F.; HILSDORF, A.W.S. 2017 First record of the Saccostrea oyster in Bertioga, São Paulo, Brazil. Boletim do Instituto de Pesca, 43(4): 638-645.
GARDUNHO, D.C.L.; GOMES, C.P.; TAGLIARO, C.H.; BEASLEY, C.R. 2012 Settlement of an unidentified oyster (Crassostrea) and other epibenthos on plastic substrates at a northern Brazilian mangrove island. Brazilian Journal of Aquatic Science and Technology, 16(1): 41-51.
HALL, T.A. 1999 BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95 âÂ"ž 98 âÂ"ž NT. Nucleic Acids Symposium Series, 41(41): 95í 98.
HEBERT, P. D.; RATNASINGHAM, S.; DE WAARD, J. R. 2003 Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London B: Biological Sciences, 270 (Suppl 1): S96-S99.
IGNACIO, B.L.; ABSHER, T.M.; LAZOSKI, C.; SOLE´-CAVA, A.M. 2000 Genetic evidence of the presence of two species of Crassostrea (Bivalvia: Ostreidae) on the coast of Brazil. Marine Biology, 136(6): 987í 991.
LAPEGUE, S.; BOUTET, I.; LEITÃO, A.; HEURTEBISI, S.; GARCIA, P.; THIRIOTUIE-VREUX, C.; BOUDRY, P. 2002 Trans-Atlantic distribution of a mangrove oyster species revealed by 16S mtDNA and karyological analyses. Biology Bull, 202(3): 232-242.
LAZOSKI, C.; GUSMÃO, J.; BOUDRY, P.; SOLE´-CAVA, A.M. 2011 Phylogeny and phylogeography of Atlantic oyster species: evolutionary history, limited genetic connectivity and isolation by distance. Marine Ecology Progress Series, 426: 197í 212.
LIBRADO, P.; ROZAS, J. 2009 DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25(11): 1451-1452.
LUDWIG, S.; PATELLA, R.; STOIEV, S.; CASTILHO-WESTPHAL, G.; GIROTTO, M. V.; OSTRENSKY, A. 2011 A molecular method to detect and identify the native species of southwestern Atlantic Crassostrea (Mollusca: Ostreidae). Zoologia (Curitiba), 28(4): 420-426.
MEDRANO, J. F.; AASEN, E.; SHARROW, L. 1990 DNA extraction from nucleated red blood cells. Biotechniques, 8(1): 43.
MELO, A. G. C. D.; VARELA, E. S.; BEASLEY, C. R.; SCHNEIDER, H.; SAMPAIO, I.; GAFFNEY, P. M.; TAGLIARO, C. H. 2010 Molecular identification, phylogeny and geographic distribution of Brazilian mangrove oysters (Crassostrea). Genetics and Molecular Biology, 33(3): 564-572.
MELO, M.A.D.; SILVA, A.R.B.; BEASLEY, C.R.; TAGLIARO, C.H. 2013 Multiplex species-specific PCR identification of native and non-native oysters (Crassostrea) in Brazil: a useful tool for application in oyster culture and stock management. Aquaculture International, 21(06): 1325í 1332.
PIE, M. R.; RIBEIRO, R. O.; BOEGER, W. A.; OSTRENSKY, A.; FALLEIROS, R. M.; ANGELO, L. 2006 A simple PCRâ€ÂRFLP method for the discrimination of native and introduced oyster species (Crassostrea brasiliana, C. rhizophorae and C. gigas; Bivalvia: Ostreidae) cultured in Southern Brazil. Aquaculture Research, 37(15): 1598-1600.
REECE, K. S.; CORDES, J. F.; STUBBS, J. B.; HUDSON, K. L.; FRANCIS, E. A. 2008 Molecular phylogenies help resolve taxonomic confusion with Asian Crassostrea oyster species. Marine Biology, 153(4): 709í 721.
RIOS, E. C. 1994 Sea shells of Brazil. 2ª ed. Rio Grande: Fundação Universidade do Rio Grande. 492p.
SAITOU, N.; NEI, M. 1987 The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution, 4(4): 406í 425.
SANGER, F.; NICKLEN, S.; COULSON, A. R. 1977 DNA sequencing with chain-terminating inhibitors. Proceedings of the national academy of sciences, 74(12): 5463-5467.
SIMONE, L. R. L.; DO AMARAL, V. S. 2014 Revision of genus Crassostrea (Bivalvia: Ostreidae) of Brazil. Journal of the Marine Biological Association of the United Kingdom, 94(4): 811-836.
SOLÉ-CAVA, A. M. 2008 Códigos de barra de DNA: o rabo que abana o cachorro. Ciência Hoje, 41(245): 65-67.
SOLÉ-CAVA, A. M.; Wí–RHEIDE, G. 2007 The perils and merits (or the Good, the Bad and the Ugly) of DNA barcoding of sponges í a controversial discussion. In: CUSTÓDIO, M.R.; LÔBO-HAJDU G.; HAJDU, E.; MURICY G. (ed.) Porifera research: biodiversity, innovation and sustainability. Série Livros (28). Rio de Janeiro: Museu Nacional. p. 603í 612.
TAMURA, K.; STECHER, G.; PETERSON, D.; FILIPSKI, A.; KUMAR, S. 2013 MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular biology and evolution, 30(12): 2725-2729.
THOMPSON, J. D.; HIGGINS, D. G.; GIBSON, T. J. 1994 Clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic acids research, 22(22): 4673-4680.
VARELA, E.S; BEASLEY, C.R.; SCHNEIDER, H.; SAMPAIO, I.; MARQUES-SILVA, N.S.; TAGLIARO, C.H. 2007 Molecular phylogeny of mangrove oysters (Crassostrea) from Brazil. Journal of molluscan studies, 73(3): 229-234.
WAKAMATSU, T. 1973 A ostra de Cananéia e o seu cultivo. São Paulo: Instituto Oceanográfico, Universidade de São Paulo. 141 p.
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2018-12-26
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